A multiplex one‐step PCR method for the simultaneous identification of Bursaphelenchus xylophilus, B. mucronatus and B. doui– three species within the xylophilus group
A multiplex one‐step PCR method for the simultaneous identification of Bursaphelenchus xylophilus, B. mucronatus and B. doui– three species within the xylophilus group
复制标题
DOI：
10.1111/j.1439-0329.2009.00638.x
复制
发表时间：
2011-02
期刊：
影响因子：
1.4
通讯作者：
K. Zhuo;M. Luo;R. Cui;J. Liao
中科院分区：
文献类型：
--
作者：
K. Zhuo;M. Luo;R. Cui;J. Liao
松材线虫，松材线虫，严重危害东亚地区的松树。拟松材线虫和双拟松材线虫在形态上与松材线虫极为相似，而且近年来在该地区频繁发现。有必要对这三个物种进行准确、快速的鉴定。在这项研究中，我们报告了一种多重一步聚合酶链式反应(PCR)的结果，该方法使用五个引物同时鉴定和区分这三个伯氏杆菌种。多重一步聚合酶链式反应在所有供试种群中扩增出一条约1000bp的片段。另外，松材线虫、拟松材线虫和豆芽孢杆菌分别扩增出约100、350和600bp的片段。这种方法简单而可靠，可以同时识别松材线虫群中的上述三个物种，这是线虫分析中经常遇到的。
Summary The pine wood nematode, Bursaphelenchus xylophilus, causes severe damage to pines in Eastern Asia. Bursaphelenchus mucronatus and B. doui resemble closely B. xylophilus morphologically, moreover they were found frequently in this area recently. It is necessary to identify the three species precisely and rapidly. In this study, we report the results of a multiplex one-step polymerase chain reaction (PCR) utilizing five primers to identify and discriminate the three Bursaphelenchus species simultaneously. The multiplex one-step PCR yielded one fragment of about 1000 bp for all Bursaphelenchus populations tested. Futhermore, B. xylophilus, B. mucronatus and B. doui produced another fragment of about 100, 350 and 600 bp respectively. This approach is simple and reliable to simultaneously identify the above three species within the xylophilus group usually encountered together in a nematode assay.